bisphosphonic acid derivatives, and useful as a bone resorption inhibitor

ABSTRACT

A bisphosphonic acid derivative of the general formula (I): ##STR1## wherein A is an optionally substituted cyclic group; R 1  is hydrogen atom or a lower alkanoyl group; R 2 , R 3 , R 4  and R 5  are the same or different and are hydrogen atom or a lower alkyl group; m is 0, 1 or 2; and n is an integer from 2 to 10, or a salt thereof is disclosed. A process for its production and a bone resorption inhibitor containing the compound of the general formula (I) or a salt thereof are also disclosed.

This application is a continuation of U.S. application Ser. No.07/718,648 filed Jun. 21, 1991, now abandoned.

FIELD OF THE INVENTION

The present invention relates to (sulfur-containingalkyl)aminomethylenebisphosphonic acid derivatives or pharmaceuticallyacceptable salts thereof which are useful as medicament having boneresorption inhibitory activity as well as anti-inflammatory activity,antirheumatic activity and the like, and a medicament containing thecompounds as an active component.

PRIOR ART

Various aminomethylenebisphosphonic acid derivatives have beensynthesized as disclosed in Japanese Patent Laid Open Publication Nos.308290/1989, 258695/1990, 184/1990, 185/1990 and the like. However, noneof them discloses (sulfur-containing alkyl)aminomethylenebisphosphonicacid derivatives of the present invention.

OBJECTS OF THE INVENTION

Although various bisphosphonic acid derivatives have been produced asbone resorption inhibitors, they are yet insufficient from the viewpointof their activity and side effects.

In view of these circumstances, the present inventors have studiedintensively to develop bisphosphonic acid derivatives which are moreuseful as bone resorption inhibitors. As a result, it has been foundthat novel bisphosphonic acid derivatives of the general formula (I) asshown hereinafter can directly effect bone to manifest excellent boneresorption inhibitory activity.

SUMMARY OF THE INVENTION

The compound of the present invention is characterized by its chemicalstructure in that it has thio, sulfinyl or sulfonyl group bound to acyclic group on the alkyl side-chain, and the compounds of the presentinvention can be used as bone resorption inhibitors.

Namely, according to the present invention, there is provided,

(1) a bisphosphonic acid derivative of the general formula (I): ##STR2##wherein A is an optionally substituted cyclic group; R¹ is hydrogen atomor a lower alkanoyl group; R², R³, R⁴ and R⁵ are the same or differentand are hydrogen atom or a lower alkyl group; m is 0, 1 or 2; and n isan integer from 2 to 10, or a salt thereof,

(2) a process for production of the compound of the general formula (I)which comprises reacting an amine derivative of the general formula(II): ##STR3## wherein all the symbols are as defined above, with anorthoformate derivative of the general formula (III):

    CH(OR.sup.6).sub.3                                         (III)

wherein R⁶ is a lower alkyl group, and a phosphite derivative of thegeneral formula (IV): ##STR4## wherein R⁷, R⁸, R⁹ and R¹⁰ are the sameor different and are a lower alkyl group, and then optionally subjectingthe resultant to acylation, oxidation and/or hydrolysis, and

(3) a bone resorption inhibitor which comprises the compound of thegeneral formula (I) or a salt thereof.

DETAILED DISCLOSURE OF THE INVENTION

In the above general formula (I), the optionally substituted cyclicgroup represented by A includes C₆₋₁₄ aromatic hydrocarbon residues suchas phenyl, naphthyl, anthryl and the like; 5 or 6-membered aromaticheterocyclic groups containing 1 to 4 hetero atoms, preferably,nitrogen, oxygen and/or sulfur atoms (e.g., pyridyl, pyrimidinyl,pyridazinyl, furyl, thienyl, imidazolyl, thiazolyl, oxazolyl isoxazolyl,thiadiazolyl, pyrazolyl, triazolyl, etc.); 5 or 6 membered aromaticheterocyclic groups containing 1 to 4 nitrogen, oxygen and/or sulfuratoms which are conjugated with a C₆₋₁₄ aromatic hydrocarbon ring orwith a 5 or 6 membered aromatic heterocyclic ring containing 1 to 4nitrogen, oxygen and/or sulfur atoms (e.g., benzothiazolyl,benzoxazolyl, benzoimidazolyl, s-triazolo[1,2-a]pyridyl,imidazo[1,2-b]pyrazinyl, indolyl, imidazo[1,2-a]pyridyl, etc.); C₃₋₇cyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl and the like; and 5 or 6-membered non-aromaticheterocyclic group containing 1 to 4 hetero atoms, preferably, nitrogen,oxygen and/or sulfur atoms (e.g., thiazolin-2-yl, imidazolin-2-yl,oxazolin-2-yl, etc.). When a cyclic group containing nitrogen atom asthe hetero atom has a substituent, such a substituent may attach toeither the carbon atom or nitrogen atom in the cyclic group.

In the above general formula (I), examples of the substituent of thering A include a halogen atom, nitro group, an optionally substitutedalkyl group, optionally substituted hydroxyl group, optionallysubstituted thiol group. These substituents may be the same or differentand the ring A may have 1 to 4, preferably 1 or 2 substituents.

The term "halogen atom" used herein includes fluorine, chlorine,bromine, iodine and the like. The alkyl group in the optionallysubstituted alkyl group is preferably, a C₁₋₇ straight or branched chainalkyl group such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl,sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl or heptyl orthe like; or a C₃₋₇ cycloalkyl group such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl or the like. They may besubstituted with 1 to 3 substituents such as a halogen atom (e.g.,fluorine, chlorine, bromine, iodine, etc.), hydroxyl group, a C₁₋₆alkoxy group (e.g., methoxy, ethoxy, propoxy, butoxy, hexyloxy, etc.)and the like.

The examples of the substituted alkyl group include trifluoromethyl,2,2,2-trifluoroethyl, trichloromethyl, hydroxymethyl, 2-hydroxyethyl,2-methoxyethyl and the like.

The substituted hydroxyl group is that having a suitable substituent,particularly, a protecting group for a hydroxyl group, for example,alkoxy, alkenyloxy, aralkyloxy, acyloxy as well as aryloxy. Examples ofthe alkoxy group include C₁₋₆ straight or branched chain alkoxy groups(e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy,sec-butoxy, tert-butoxy, pentoxy, isopentoxy, neopentoxy, hexyloxy,etc.) and C₄₋₆ cycloalkoxy groups (e.g., cyclobutoxy, cyclopentoxy,cyclohexyloxy, etc.) and examples of the alkenyloxy group preferablyinclude C₂₋₆ alkenyloxy groups such as allyloxy, crotyloxy,2-pentenyloxy, 3-hexenyloxy, 2-cyclopentenylmethoxy and the like. As thearalkyloxy group, preferably, C₆₋₁₉ aralkyloxy groups, more preferably,C₆₋₁₄ aryl-C₁₋₄ alkyloxy groups (e.g., benzyloxy, phenethyloxy, etc.)may be used. As the acyloxy group, preferably, alkanoyloxy groups, forexample, C₂₋₇ alkanoyloxy groups (e.g., acetyloxy, propionyloxy,n-butyryloxy, iso-butyryloxy, hexanoyloxy, etc.) may be used. As thearyloxy group, preferably, C₆₋₁₄ aryloxy groups (e.g., phenoxy,biphenyloxy, etc.) may be used. These groups may be further substitutedwith 1 to 3 substituents such as the above-described halogen atom,hydroxyl group, C₁₋₆ alkoxy group and the like. Examples of thesubstituted hydroxyl group include trifluromethoxy, difluoromethoxy,2,2,2-trifluoroethoxy, 2-methoxyethoxy, 4-chlorobenzyloxy,2-(3,4-dimethoxyphenyl)ethoxy and the like.

The substituted thiol group is a thiol group having a suitablesubstituent, particularly, a protecting group for thiol group, forexample, alkylthio, aralkylthio, acylthio. As the alkylthio group,preferably a C₁₋₆ straight or branched chain alkylthio group (e.g.,methylthio, ethylthio, propylthio, isopropylthio, butylthio,isobutylthio, sec-butylthio, tert-butylthio, pentylthio, isopentythio,neopentylthio, hexylthio, etc.), and a C₄₋₇ cycloalkylthio (e.g.,cyclobutylthio, cyclopentylthio, cyclohexylthio, cycloheptylthio, etc.)may be used. As the aralkylthio group, preferably, a C₇₋₁₉ aralkylthiogroup, more preferably a C₆₋₁₄ aryl-C₁₋₄ alkylthio group such asbenzylthio or phenetylthio may be used. As the acylthio group,preferably, alkanoylthio group such as C₂₋₇ alkanoylthio (e.g.,acetylthio, propionylthio, n-butyrylthio, iso-butyrylthio, hexanoylthio,etc.) may be used. These groups may be further substituted with, forexample, 1 to 3 substituents such as the above-described halogen atom,hydroxyl group, C₁₋₆ alkoxy group and the like. Examples of thesubstituted thiol group include trifluoromethylthio, difluoromethylthio,2,2-trifluoroethylthio, 2-methoxyethylthio, 4-chlorobenzylthio,3,4-dichlorobenzylthio, 4-fluorobenzylthio,2-(3,4-dimethoxyphenyl)ethylthio and the like.

Examples of the substituted aromatic hydrocarbon group include4-chlorophenyl, 2-fluorophenyl, 4-nitorophenyl, 3-methylphenyl,3-trifluoromethylphenyl, 5,6,7,8-tetrahydro-2-naphthyl, 4-methoxyphenyl,3,4-methylenedioxyphenyl, 4-(4-chlorbenzyloxy)phenyl, 4-acetoxyphenyl,3-methylthiophenyl and the like.

Examples of the substituted aromatic heterocyclic group include2-chloro-4-pyridyl, 5-nitro-2-pyridyl, 3-hydroxy-2-pyridyl,6-methoxy-2-pyridyl, 2-methyl-4-pyridyl, 4-methyl-2-pyrimidinyl,4-hydroxy-6-methyl-2-pyrimidinyl, 5-trifluoromethyl-2-benzothiazolyl andthe like.

Examples of the substituted heterocyclic group include5-phenyl-2-thiazolin-2-yl, 5-methyl-2-oxazoiin-2-yl,1-methyl-2-imidazolin-2-yl and the like.

As the lower alkanoyl group represented by R¹, C₁₋₆ alkyl-carbonyl group(e.g., acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl,pivaloyl, hexanoyl, cyclopentanecarbonyl, etc.) may be used. Preferably,R¹ is, for example, acetyl, propionyl, butyryl or the like.

The lower alkyl group represented by R², R³, R⁴ and R⁵ include C₁₋₄straight or branched chain alkyl groups, for example, methyl, ethyl,propyl, butyl, isopropyl, isobutyl, tert-butyl and the like.

Among the above groups, the ring A is preferably benzene ring, pyridinering or pyrimidine ring which may be substituted with a halogen atom, analkyl group or an alkoxy group.

The suitable salts of the compound (I) are pharmaceutically acceptablesalts, for example, inorganic salts such as alkali metal salts (e.g.,sodium salt, potassium salt, etc.), alkaline earth metal salts (e.g.,calcium salt, magnesium salt, etc.) or ammonium salt; salts with organicbases such as methylamine salt, ethylamine salt, propylamine salt,isopropylamine salt, butylamine salt, tert-butylamine salt,dimethylamine salt, diethylamine salt, trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, dicyclohexylaminesalt, N,N'-dibenzylethylenediamine salt and the like; organic acid saltssuch as formate, acetate, trifluoroacetate, maleate, tartrate,methanesulfonate, benzenesulfonate, toluenesulfonate and the like;inorganic acid salts such as hydrochloride, hydrobromide, sulfonate andthe like; and salts with amino acids such as lysine, glutamic acid andthe like.

The compound (I) or a salt thereof can be produced by the known methods.

Namely, the compound (I) or its salt can be produced by reacting anamine derivative of the general formula ( II ): ##STR5## wherein A is anoptionally substituted cyclic group; and n is an integer of 2 to 10,with an orthoformate derivative of the general formula (III):

    CH(OR.sup.6).sub.3                                         (III)

wherein R⁶ is a lower alkyl group, and a phosphite derivative of thegeneral formula (IV): ##STR6## wherein R⁷, R⁸, R⁹ and R¹⁰ are the sameor different and are a lower alkyl group, and then optionally subjectingthe resultant to acylation, oxidation and/or hydrolysis.

For example, the compound (I) or a salt thereof can be produced by thefollowing methods. The salts of the compounds described hereinafter maybe the same as those described with respect to the compound (I).##STR7## wherein all the symbols are as defined above. ##STR8## whereinR^(1') represents a lower acyl group, and the other symbols are asdefined above. ##STR9## wherein k represents 1 or 2, and the othersymbols are as defined above. ##STR10## wherein all the symbols are asdefined above.

Method D-2

The production of a bisphosphonic acid diester.

Method D-3

The production of a bisphosphonic acid mono- or triester.

Each method is illustrated in detail below.

Method A

This method produces the bisphosphonate derivative (I-1) by reacting theamine derivative (II) with the orthoformate derivative (III) and thephosphite derivative (IV) in amounts suitable for the reaction. Thereaction is usually conducted at 80° C. to 200° C., preferably at 100°C. to 170° C. for 10 minutes to 24 hours.

Method B

In this method, the compound (I-2) is produced by acylation of thecompound (1-I) produced by the method A. This acylation is conducted byreacting the compound (1-I) with 1 to 2 equivalents of an acylatingagent (acid anhydride, acid halide, etc.) in a solvent or without anysolvent. As the solvent, benzene, xylene, toluene, chloroform,dichloromethane, ethyl acetate, ether, tetrahydrofuran or the like maybe used. The reaction is conducted at 0° C. to 100° C. for 30 minutes to10 hours.

Method C

This method is conducted by oxidation using an oxidizing agent accordingto the conventional method. Such an oxidizing agent is a mild one whichproduces less substantial effect on the skeleton of thesulfur-containing heterocyclic compound, and preferably,m-chloroperbenzoic acid, hydrogen peroxide, peresters, sodiummetaperiodate or the like may be used.

This reaction is conducted in an organic solvent which has no adverseeffect on the reaction.

As the solvent, for example, a halogenated hydrocarbon (e.g., methylenechloride, chloroform, dichloroethane, etc.), or hydrocarbon (e.g.,benzene, toluene, etc. ) or a mixed solvent thereof may be used.

When equimolar or less of the oxidizing agent is used based on thecompound (I-1) or (I-2), the compound (I-3) wherein k is 1 ispreferentially produced. When more than equimolar amount of theoxidizing agent is used, the compound (I-3) wherein k is 1 is furtheroxidized to produce the compound (I-3) wherein k is 2.

This reaction proceeds at a temperature of not higher than roomtemperature (20° C. to 30° C.). Preferably, the reaction temperature isbetween about -50° C. to 20° C.

The reaction time is from 30 minutes to 10 hours.

Method D-1

In this method, the bisphosphonates (i-1), (I-2) and (I-3) produced bythe above methods A to C are subjected to hydrolysis to produce thecorresponding bisphosphonic acid (I-4).

This reaction is conducted using an inorganic acid such as hydrochloricacid, hydrobromic acid or the like or a halogenated trialkylsilane in asolvent which has no adverse effect on the reaction. When an inorganicacid such as hydrochloric acid, hydrobromic acid or the like is used, analcohol such as methanol, ethanol, 2-methoxyethanol, ethylene glycol,propanol, butanol or th like, water or a mixed-solvent thereof is usedas the solvent. The amount of the acid used is usually large excess andthe reaction temperature is 0° C. to 150° C., preferably 30° C. to 100°C., and the reaction time is 1 to 50 hours.

When a halogenated alkylsilane such as chlorotrimethylsilane,bromotrimethylsilane, iodotrimethylsilane or the like is used, ahalogenated hydrocarbon such as carbon tetrachloride, chloroform,dichloromethane, 1,2-dichloroethane, 1,1,2,2-tetrachloroetahne,acetonitrile or the like or a mixed solvent thereof may be used as thesolvent.

The amount of the halogenated alkylsilane used is 4 to 10 equivalent,preferably 5 to 8 equivalent of the compound (I-1), (I-2) or (I-3). Thereaction temperature is -30° C. to 100° C., preferably -10° C. to 50° C.and the reaction time is 30 minutes to 100 hours.

In order to convert the bisphosphonic acid thus obtained into a saltthereof, the acid is treated according to the conventional method usinga base such as potassium hydroxide, sodium hydroxide, sodium methoxide,ammonia, organic amines or the like.

Method D-2

In this method, the bisphosphonic acid tetraester (I-4) produced by themethod A is subjected to hydrolysis with a base to produce thebisphosphonic acid diester.

The amount of the base (e.g., sodium hydroxide, potassium hydroxide,etc.) to be used is 2 to 2.2 molar equivalent based on the compound(I-1) and the reaction is conducted in a solvent containing wateraccording to the conventional method.

Method D-3

In this method, the bisphosphonic acid tetraesters (I-1), (I-2) and(I-3) produced by the methods A to C are subjected to partial hydrolysiswith a halogenated alkylsilane to produce the bisphosphonic acidtetraesters.

The amount of the halogenated alkylsilicon (e.g., chlorotrimethylsilane,bromotrimethylsilane, iodotrimethylsilane, etc.) to be used is 1 to 1.2molar equivalent based on the compound (I-1), (I-2) or (I-3) in the caseof the triester production, and 3 to 3.3 molar equivalent based on thecompound in the case of the monoester production, and the reaction isconducted according to the method D-1.

The bisphosphonic acid derivative (I) thus obtained can be isolated andpurified according to known means for separation and purification, forexample, by concentration, concentration under reduced pressure, solventextraction, precipitation, recrystallization, chromatography and thelike.

The starting compound (II) of the present invention can be produced, forexample, by the following method. ##STR11## wherein the ring A and n areas defined above, and X represents a leaving group.

In this method, the compound (V) is reacted with the compound (VI) inthe presence of a base to produce the compound (VII). The leaving grouprepresented by X is, for example, a halogen, preferably chlorine,bromine or iodine, or an activated hydroxyl group, for example, anesterified organic sulfonic acid residue (e.g., p-toluenensulfonyloxygroup, etc.), a C₁₋₄ alkylsulfonyloxy group (e.g., methanesulfonyloxygroup, etc.) or an esterified organic phosphoric acid residues such asdiphenylphosphoryloxy group, dibenzylphosphoryloxy group,dimethylphosphoryloxy group and the like. The reaction of the compounds(V) with (VI) is conducted in a suitable solvent. Examples of thesolvent include aromatic hydrocarbons such as benzene, toluene, xylene;ethers such as dioxane, tetrahydrofuran, dimethoxyethane; alcohols suchas methanol, ethanol, propanol; ethyl acetate; acetonitrile; pyridine;N,N-dimethylformamide; dimethylsulfoxide; chloroform; dichloromethane;1,2-dichloroethane; 1,1,2,2-tetrachloroethane; acetone; 2-butanone and amixed solvent thereof. The reaction of the compounds (V) with (VI) isconducted in the presence of a suitable base, for example, an alkalimetal salt such as sodium hydroxide, potassium hydroxide, potassiumcarbonate, sodium carbonate, sodium bicarbionate or the like; or anamine such as pyridine, N,N-dimethylaniline or the like. The amount ofthe base to be used is preferably about 1 to 5 moles based on thecompound (V). This reaction is usually conducted at -20° C. to 150° C.,preferably about 0° C. to 130° C. for 1 to 10 hours. ##STR12## whereinthe ring A and n are as defined above.

In this method, the compound (V) is firstly reacted with about equimolaramount of the compound (VIII) in the presence of a base to produce thecompound (IX). The reaction between the compounds (V) and (VIII) isconducted in the same manner as that of Method E. ##STR13## wherein thering A and n are as defined above.

In this method, the compound (IX) obtained in the first step of Method Fis reacted with about equimolar amount of potassium phthalimide toproduce the compound (VII). The reaction between the compound (IX) andpotassium phthalimide is conducted in a suitable solvent. Examples ofthe solvent include aromatic hydrocarbons such as benzene, toluene,xylene and the like; ethers such as dioxane, tetrahydrofuran,dimethoxyethane and the like; alcohols such as methanol, ethanol,propanol and the like; ethyl acetate; acetonitrile; pyridine;N,N-dimethylformamide; dimethylsulfoxide; chloroform; dichloromethane;1,2-dichtoroethane; 1,1,2,2-tetrachloroethane; acetone; 2-butanone and amixed solvent thereof. This reaction is usually conducted at -20° C. to150° C., preferably about 30° C. to 130° C. for 1 to 10 hours. ##STR14##wherein the ring A and n are as defined above.

In this method, the compound (VII) prepared by the methods E and F isreacted with hydrazine hydrate to produce the compound represented bythe general formula (II). The reaction between the compound (VII) andhydrazine hydrate is conducted in a suitable solvent. Examples of thesolvent include aromatic hydrocarbons such as benzene, toluene, xyleneand the like; ethers such as dioxane, tetrahydrofuran, dimethoxyethaneand the like: alcohols such as methanol, ethanol, propanol and the 1 ike; N,N-dimethylformamide; dimethylsulfoxide; chloroform;dichloromethane; 1,2-dichloroethane; 1,1,2,2-tetrachloroethane and amixed solvent thereof. The amount of the hydrazine hydrate used is 1 to10 molar equivalent, preferably, 1.2 to 5 molar equivalent based on thecompound (VII). This reaction is usually conducted at -20° C. to 150°C., preferably about 0° C. to 100° C. for 1 to 10 hours.

The compound (I) or a salt thereof provided by the present invention hasbone resorption inhibitory activity and prevent bone loss caused by boneresorption.

Accordingly, the compound of the present invention can be used forprevention or treatment of osteoporosis of mammals (e.g., mouse, rat,rabbit, dog, cat, cattle, pig, man, etc. ).

When the compound of the present invention is administered to man, itcan be administered by way of either oral or parenteral route. Acomposition for oral administration may be a solid or liquid dosageforth, for example, tablets (including sugar coated tablets, film coatedtablets ), pills, granules, powder, capsules (including soft capsules),syrup, emulsion, suspension and the like. Such compositions can beprepared by known methods and may contain a carrier or excipient whichis conventionally used in the field of pharmaceutical preparations. Forexample, a carrier or excipient for tablets includes lactose, starch,sucrose, magnesium stearate and the like.

The composition for parenteral administration may be an injectionpreparation or suppository, and such an injection preparation includesdosage forms for subcutaneous injection, intracutaneous injection,intramuscuiar injection and the like. The injection preparation can beprepared by a known method, namely, by a method wherein the compound (i)is suspended or emulsified in a sterilized aqueous or oily solutionwhich is usually used for injection preparations. Such an aqueoussolution for injection includes physiological saline, isotonic solutionand the like. If desired, the solution may be used in a combination witha suitable suspending agent, for example, sodium carboxymethylcellulose,a non-ionic surfactant or the like. The oily solution includes sesameoil, soybean oil and the like, and may be used in a combination with asolubilizing agent such as benzyl benzoate, benzyl alcohol, or the like.The injection preparation thus obtained is usually filled in a suitableampoule.

When the compound (I) or a salt thereof is used as medicine fortreatment of osteoporosis, the daily dosage for an adult is 1 to 500 mg,preferably 10 to 200 mg in the case of oral administration.

The method for measurement of bone resorption inhibitory activity andeffect of osteoporosis treatment of the compound (I) as well as theresult thereof are set forth below.

Bone resorption inhibitory activity

Measurement of bone resorption inhibitory activity was conductedaccording to Raisz's method [J. Clin. Invest., 44, 103-116 (1965)].Namely, a rat (Sprague-Dawley, 19th day of pregnancy) was subcutaneouslyinjected with 50 μCi 45_(Ca) (calcium isotope, CaCl₂ solution) andsubjected to abdominal incision on the next day to aseptically removefetal rats. Both cubital bones (radius, cubitus) were cut off underanatomic microscopic observations and the connective tissue andcartilage were removed as much as possible to obtain a sample for a boneculture. Each piece of the bones was cultivated in 0.6 ml BGJ_(b) medium(Fitton-Jackson modification [GIBCO Laboratories (U.S.A.)] containing 2mg/ml of bovine serum albumin) at 37° C. for 24 hours, and then thecompound to be tested was added so that the concentration thereof became10 μg/ml. Then, the cultivation was further continued in the abovemedium for 2 days, and the radioactivity of ⁴⁵ Ca in the medium and thatin the bone were measured. The amount (%) of ⁴⁵ Ca released in themedium from the bone was calculated by the following equation. ##EQU1##

The bone obtained from the litter fetus and cultivated according to thesame manner without addition of a compound to be tested for 2 days wasused as a control group. The average value±standard deviation of thedata obtained from five bones of each group was calculated. The ratio(%) of the value thus obtained to that of the control group was obtainedand shown in Table 1.

                  TABLE 1                                                         ______________________________________                                                   Release of Ca.sup.45                                               Ex. No.    (%, to the control group)                                          ______________________________________                                        5          73                                                                 ______________________________________                                    

Effect on treatment of osteoporosis

The both ovaries of a SAM-R/1 mouse (13-weeks old) were removed and atest compound was orally administered to the animal for 6 days per oneweek, totally for 17 days in 3 weeks from the next day of the operation.On the next day of the last administration, the left femur of the mousewas removed. Trochlea was removed from the femur, and then the distalone-third of the femur was cut at right angles to the longitudinal axis.The bone-marrow was removed by dissolving with 0.2N aqueous potassiumhydroxide, and placed in a glass tube. It was placed in an electricdryer and dried at 100° C. for 3 hours, then dry weight was measured.

The average value±standard deviation of the data obtained from themeasurement of 6 to 8 mice per group are shown in Table 2.

                  TABLE 2                                                         ______________________________________                                                      Daily Dose                                                                              Weight (Dry Basis)                                    Group         (mg/kg)   (mg)                                                  ______________________________________                                        Sham operation                                                                              0         10.18                                                 control group           ±0.18**                                            Ovary removed 0         9.16                                                  control group           ±0.09                                              Ovary removed 30        10.31                                                 compound (Ex. 5)        ±0.31**                                            administered                                                                  ______________________________________                                         Significance with respect to the ovary removed control group                  *: p < 0.05, **: p < 0.01                                                

Effect on prevention and treatment of osteoporosis

A male Sprague-Dawley rat (6-weeks old) was administered with thespecimen (the compound) intraperitoneally for 2 days, and on the thirdday, the right sciatic nerve of the rat was cut out. Both cnemis wereremoved on the 17th day. The proximal half portion of the cnemis was cutout at right angles to the longitudinal axis and then dried at 110° C.for 6 hours. The dry weight was measured.

The average value±standard deviation obtained from of the data from themeasurement of 6 mice per group are shown in Tables 3 and 4.

                  TABLE 3                                                         ______________________________________                                                  Daily Dose                                                                             Weight (Dry Basis) (mg)                                    Group       (mg/kg)    Right cnemis                                                                             Left cnemis                                 ______________________________________                                        Sham operation                                                                            0           99.8 ± 3.9**                                                                         109.7 ± 1.5                              control group                                                                 Operation   0          79.4 ± 2.1                                                                            106.7 ± 1.1                              control group                                                                 Operation   1          146.8 ± 5.8**                                                                          157.0 ±                                 compound (Ex. 10)                 6.6**                                       administered group                                                            ______________________________________                                         Significance with respect to the operation control group                      **: p < 0.01                                                             

                  TABLE 4                                                         ______________________________________                                                  Daily Dose                                                                             Weight (Dry Basis) (mg)                                    Group       (mg/kg)    Right cnemis                                                                             Left cnemis                                 ______________________________________                                        Sham operation                                                                            0           98.1 ± 3.5**                                                                         97.5 ± 0.6                               control group                                                                 Operation   0          77.3 ± 1.7                                                                            99.6 ± 2.4                               control group                                                                 Operation   1          134.4 ± 4.5**                                                                          143.7 ± 5.2**                           compound (Ex. 41)                                                             administered group                                                            ______________________________________                                         Significance with respect to the operation control group                      **: p < 0.01                                                             

The following Reference Examples and Examples further illustrate thepresent invention in detail but are not to be construed to limit thescope thereof. Reference Example 1

A mixture of 2-mercaptopyrimidine (7.3 g), N-(2-bromoethyl)phthalimide(16.5 g), potassium carbonate (10.8 g) and N,N-dimethylformamide (DMF)(85 ml) was stirred at room temperature for 3 hours. The reactionmixture was poured into water and the crystals separated were collectedby filtration to obtain N-[2-(2-pyrimidinylthio)ethyl]-phthalimide (17.3g, 93%). The resultant was recrystallized from ethanol-isopropyl etherto obtain pale yellow prisms, m.p. 149°-150° C.

Reference Examples 2 to 20

According to the same manner as that described in Reference Example 1,the compounds shown in Tables 5 and 6 were obtained.

                  TABLE 5                                                         ______________________________________                                         ##STR15##                                                                     Ex.Ref.                                                                            ##STR16##   n     (%)Yield                                                                            (°C.)m.p.                                                                     Recrystn. solvent                        ______________________________________                                              ##STR17##  2     78    53.54  ether-hexane                              3                                                                                   ##STR18##  3     88    83-84  isopropyl ether                           4                                                                                   ##STR19##  4     83    65-66  isopropyl ether                           5                                                                                   ##STR20##  4     85    84-85  isopropyl ether                           6                                                                                   ##STR21##  2     90    98-99  isopropyl ether                           7                                                                                   ##STR22##  3     88    103-104                                                                              acetone- isopropyl ether                  8                                                                                   ##STR23##  4     99    crude*.sup.1)  oil                                                                    --                                       9                                                                                   ##STR24##  2     88    147-148                                                                              ethyl acetate                             10                                                                                  ##STR25##  3     90    131-132                                                                              ethyl acetate                             11                                                                                  ##STR26##  4     94    105-106                                                                              isopropyl ether                           12                                                                                  ##STR27##  2     76    108-109                                                                              ethyl acetate hexane                      ______________________________________                                    

                  TABLE 6                                                         ______________________________________                                         ##STR28##                                                                     Ex.Ref.                                                                            ##STR29##     n     (%)Yield                                                                            (°C.)m.p.                                                                   Recrystn. solvent                        ______________________________________                                        13                                                                                  ##STR30##    4     89    43- 45                                                                             isopropyl ether- hexane                   14                                                                                  ##STR31##    2     92    127- 128                                                                           ethanol                                   15                                                                                  ##STR32##    4     98    100- 101                                                                           ethanol                                   16                                                                                  ##STR33##    3     85    95- 96                                                                             ethanol- isopropyl ether                  17                                                                                  ##STR34##    4     98    100- 101                                                                           ethyl acetate- hexane                     18                                                                                  ##STR35##    4     78    174- 176                                                                           ethyl acetate                             19                                                                                  ##STR36##    4     97    84- 85                                                                             ethyl acetate- hexane                     20                                                                                  ##STR37##    4     73    127- 128                                                                           ethyl acetate                             ______________________________________                                         *.sup.1)NMR(δppm CDCl.sub.3): 1.7-1.95(4H, m), 3.22(2H, t, J=7Hz),      3.73(2H, t, J=7Hz), 6.96(1H, ddd, J=7, 5,1Hz), 7.15(1H, ddd, J=8.1, 1Hz),     7.46(1H, ddd, J=8, 7, 2Hz), 7.7-7.75(2H, m), 7.8-7.85(2H, m), 8.40(1H,        ddd, J=5, 2, 1Hz).                                                       

Reference Example 21

A mixture of 1-bromo-5-chloropentane (10.0 g), thiophenol (5.94 g),potassium carbonate (7.45 g) and N,N-dimethylformamide (DMF) (50 ml) wasstirred at room temperature for 4 hours. The reaction mixture was pouredinto water and extracted with ethyl acetate. The ethyl acetate layer waswashed with water, dried (MgSO₄), and then concentrated to dryness.Potassium phthalimide (11.0 g) and N,N-dimethylformamide (100 ml) wereadded to the residue and the resultant was stirred at 90° C. for 2hours. The reaction mixture was poured into water and the mixture wasextracted with ethyl acetate. The ethyl acetate layer was washed withwater, dried (MgSO₄), and then concentrated to dryness. The crystalswere collected by filtration and recrystallized from isopropyl ether toobtain N-(5-phenylthiopentyl)phthalimide (14.9 g, 85%) as colorlessprisms, m.p. 87°-88° C.

Reference Examples 22 to 26

According to the same manner as that described in Reference Example 21,the compounds shown in Table 7 were obtained.

                  TABLE 7                                                         ______________________________________                                         ##STR38##                                                                     Ex.Ref.                                                                            ##STR39##  n     (%)Yield                                                                            m.p. (°C.)                                                                     Recrystn, solvent                        ______________________________________                                        22                                                                                  ##STR40## 6     91    51-52   isopropyl ether- hexane                   23                                                                                  ##STR41## 5     73    64-65   isopropyl ether                           24                                                                                  ##STR42## 6     91    crude*.sup.1)  oil                                                                     --                                       25                                                                                  ##STR43## 5     79    90-91   ethyl acetate- hexane                     26                                                                                  ##STR44## 5     87    95-96   ethyl acetate- hexane                     ______________________________________                                         *.sup.1 NMR(δppm in CDCl.sub.3):1.25-1.8(8H, m), 3.15(2H, t, J=7Hz)     3.68 (2H, t, J=7Hz), 6.95(1H, ddd, J=7, 5, 1Hz), 7.15(1H, ddd, J=8, 1,        1Hz), 7.46(1H, ddd, J=8, 7, 2Hz), 7.65-7.75(2H, m), 7.8-7.9(2H, m),           8.41(1H, ddd, J=5, 2, 1Hz).                                              

Reference Example 27

A mixture of N-[2-(2-pyrimidinylthio)ethyl]phthalimide (17.1 g),hydrazine hydrate (21 g) and ethanol (200 ml) was stirred for one hourunder reflux. The crystals separated was filtered off, and the filtratewas concentrated under reduced pressure to obtain2-(2-pyrimidinylthio)ethylamine (6.6 g 71%) as an oil.

NMR (δ ppm in CDCl₃): 1.61 (2H, s), 3.04 (2H, t, J=6Hz), 3.27 (2H, t,J=6Hz), 6.98 (1H, t, J=5 Hz), 8.52 (2H, d, J=5 Hz).

Reference Examples 28 to 52

According to the same manner as that described in Reference Example27,the compounds shown in Tables 8 to 11 were obtained as oil.

                                      TABLE 8                                     __________________________________________________________________________     ##STR45##                                                                     Ref. Ex.                                                                           ##STR46##                                                                             n                                                                               Yield (%)                                                                           NMR (δppm in CDCl.sub.3)                          __________________________________________________________________________    28                                                                                  ##STR47##                                                                            2 87    1.57(2H, s), 2.9-3.1(4H, m), 7.2-7.4(5H, m)              29                                                                                  ##STR48##                                                                            3 80    1.42(2H, s), 1.7-1.9(2H, m), 2.83(2H, t, J=7Hz),                              2.99(2H, t, J=7Hz), 7.15-7.4(5H, m)                      30                                                                                  ##STR49##                                                                            4 99    1.29(2H, s), 1.5-1.8(4H, m), 2.71(2H, t, J=7Hz),                              2.94(2H, t, J=7Hz), 7.1-7.4(5H, m)                       31                                                                                  ##STR50##                                                                            5 94    1.4-1.8(8H, m), 2.69(2H, t, J=7Hz), 2.93 (2H, t,                              J=7Hz), 7.1-7.4(5H, m)                                   32                                                                                  ##STR51##                                                                            6 92    1.25-1.75(10H, m), 2.68(2H, t, J=7Hz), 2.92(2H, t,                            J=7Hz), 7.1-7.4(5H, m)                                   33                                                                                  ##STR52##                                                                            4 99    1.5-1.75(6H, m), 2.71(2H, t, J=7Hz), 2.91 (2H, t,                             J=7Hz), 7.25(4H, s)                                      34                                                                                  ##STR53##                                                                            2 75    1.63(2H, s), 3.00(2H, t, J=7Hz), 3.29(2H, t, J=7Hz),                          6.97(1H, ddd, J=7, 5, 1Hz), 7.19(1H, ddd, J=8, 1,                             1Hz), 7.47(1H, ddd, J=8, 7, 2Hz), 8.41(1H, ddd, J=5,                          2, 1Hz)                                                  35                                                                                  ##STR54##                                                                            3 82    1.57(2H, s), 1.8-1.95(2H, m), 2.83(2H, t, J= 7Hz),                            3.25(2H, t, J=7Hz), 6.97(1H, ddd, J=7, 5, 1Hz),                               7.17(1H, ddd, J=8, 1, 1Hz), 7.47(1H, ddd, J=8, 7,                             2Hz), 8.42(1H, ddd, J=5, 2, 1Hz)                         __________________________________________________________________________

                                      TABLE 9                                     __________________________________________________________________________     ##STR55##                                                                     Ref. Ex.                                                                           ##STR56##                                                                            n                                                                               Yield (%)                                                                           NMR (δppm in CDCl.sub.3)                           __________________________________________________________________________    36                                                                                  ##STR57##                                                                           4 78    1.36(2H, s), 1.5-1.85(4H, m), 2.74 (2H, t, J=7Hz),                            3.18(2H, t, J=7Hz), 6.97(1H, ddd, J= 7, 5, 1Hz),                              7.17(1H, ddd, J=8, 1, 1Hz), 7.47 (1H, ddd, J=8, 7,                            2Hz), 8.42(1H, ddd, J=5, 2, 1Hz)                          37                                                                                  ##STR58##                                                                           5 91    1.3-1.65(6H, m), 1.65-1.85(2H, m), 2.69 (2H, t,                               J=7Hz), 3.17(2H, t, J=7Hz), 6.96(1H, ddd, J=7, 5,                             1Hz), 7.16(1H, ddd, J=8, 1, 1Hz), 7.46(1H, ddd, J=8,                          7, 2Hz), 8.41(1H, ddd, J=5, 2, 1Hz)                       38                                                                                  ##STR59##                                                                           6 95    1.3-1.65(8H, m), 1.65-1.8(2H, m), 2.68(2H, t, J=7Hz),                         3.16(2H, t, J=7Hz), 6.96(1H, ddd, J=7, 5, 1Hz),                               7.16(1H, ddd, J=8, 1, 1Hz), 7.46(1H, ddd, J=8, 7,                             2Hz), 8.41(1H, ddd, J=5, 2, 1Hz)                          39                                                                                  ##STR60##                                                                           2 94    1.6-1.9(2H, broad), 3.0-3.2(4H, m), 7.13 (2H, d,                              J=6Hz), 8.39(2H, d, J=6Hz)                                40                                                                                  ##STR61##                                                                           3 97    1.57(2H, s), 1.8-1.95(2H, m), 2.87(2H, t, J=7Hz),                             3.06(2H, t, J=7Hz), 7.12(2H, d, J= 6Hz), 8.38(2H, d,                          J=6Hz)                                                    41                                                                                  ##STR62##                                                                           4 99    1.50-1.85(6H, m), 2.76(2H, t, J=7Hz), 2.98 (2H, t,                            J=7Hz), 7.10(2H, d, J=6Hz), 8.37 (2H, d, J=6Hz)           42                                                                                  ##STR63##                                                                           5 90    1.4-1.6(6H, m), 1.6-1.8(2H, m), 2.6-2.8 (2H, m),                              2.98(2H, t, J=7Hz), 7.10(2H, d, J=6 Hz), 8.38(2H, d,                          J=6Hz)                                                    __________________________________________________________________________

                                      TABLE 10                                    __________________________________________________________________________     ##STR64##                                                                     Ref. Ex.                                                                           ##STR65##                                                                               n                                                                               Yield (%)                                                                           NMR (δppm in CDCl.sub.3)                        __________________________________________________________________________    43                                                                                  ##STR66##                                                                              4 89    1.57-1.72(6H, m), 2.72(2H, t, J=7Hz), 3.07 (2H, t,                            J=7Hz), 3.62(3H, s), 6.92(1H, d, J= 1Hz), 7.05(1H,                            d, J=1Hz)                                              44                                                                                  ##STR67##                                                                              2 93    1.63(2H, s), 2.96(2H, double t, J=6and1 Hz),                                  3.13(2H, double t, J=6 and 1Hz), 3.64(3H, s),                                 6.93(1H, d, J=1Hz), 7.05 (1H, d, J=1Hz)                45                                                                                  ##STR68##                                                                              4 83    1.49(2H, s), 1.56-1.67(2H, m), 1.78-1.93 (2H, m),                             2.72(3H, s), 2.75(2H, t, J=7Hz), 3.32(2H, t,                                  J=7Hz)                                                 46                                                                                  ##STR69##                                                                              2 79    1.61(2H, s), 2.73(3H, s), 3.10(2H, t, J= 7Hz),                                3.40(2H, t, J=6Hz)                                     47                                                                                  ##STR70##                                                                              5 88    1.43-1.53(6H, m), 1.73-1.80(2H, m), 2.71(2H, t,                               J=7Hz), 3.16(2H, t, J=7Hz), 6.95(1H, t, J=5Hz),                               8.51(2H, d, J=5Hz)                                     48                                                                                  ##STR71##                                                                              4 87    1.40(2H, s), 1.57-1.84(4H, m), 2.75(2H, t, J=7Hz),                            3.17(2H, t, J=7Hz), 6.95(1H, t, J= 5Hz), 8.51(2H,                             d, J=5Hz)                                              49                                                                                  ##STR72##                                                                              3 77    1.57(2H, s), 1.9(2H, m), 2.86(2H, t, J=7Hz),                                  3.23(2H, t, J=7Hz), 6.96(1H, t, J=5Hz), 8.51(2H,                              d, J=5Hz)                                              __________________________________________________________________________

                                      TABLE 11                                    __________________________________________________________________________     ##STR73##                                                                     Ref. Ex.                                                                           ##STR74##                                                                              n                                                                               Yield (%)                                                                           NMR (δppm in CDCl.sub.3)                         __________________________________________________________________________    50                                                                                  ##STR75##                                                                             4 77    1.48-1.80(6H, m), 2.72(2H, t, J=7Hz), 3.12 (2H, t,                            J=7Hz), 3.38(2H, t, 8Hz), 4.21 (2H, t, J=8Hz)           51                                                                                  ##STR76##                                                                             4 96    1.5-2.0(6H, m), 2.72(2H, t, J=7Hz), 3.17(2H, t,                               3=7Hz), 6.92(1H, t, J=7Hz), 7.31 (1H, t, J=7Hz),                              7.77(1H, d, J=7Hz), 8.12(1H, d, J=7Hz)                  52                                                                                  ##STR77##                                                                             4 99    1.55-1.95(6H, m), 2.77(2H, t, J=7Hz), 2.93(2H, t,                             J=7Hz), 7.0-7.3(3H, m), 8.27 (1H, ddd, J=5, 2,          __________________________________________________________________________                          1Hz)                                                

Reference Example 53

According to the same manner as that described in Reference Example 21,N-[4-[(4-methoxyphenyl)thio]butyl]phthalimide was obtained andrecrystallized from acetoneisopropyl ether, m.p. 61°-62° C.

Reference Example 54

According to the same manner as that described in Reference Example1,N-[4-[(2-thiazolyl)thio]butyl]phthalimide was obtained andrecrystallized from ethanol, m.p. 60°-61° C.

Reference Example 55

According to the same manner as that described in Reference Example 1,N-[4-[(1-methyl-1, 2, 3, 4-tetrazol-5-yl)thio]butyl]phthalimide wasobtained and recrystallized from ethanol, m.p. 92°-93° C.

Reference Example 56

According to the same manner as that described in Reference Example 27,4-[(4-methoxyphenyl)thio]butylamine was obtained as an oil.

NMR (δ ppm in CDCl₃): 1.50 (2H, s), 1.53-1.88 (4H, m), 2.77 (2H, t, J=7Hz), 3.77 (3H, s), 3.93 (2H, t, J=7 Hz), 6.83 (4H, s).

Reference Example 57

According to the same manner as that described in Reference Example 27,4-[(4-methoxyphenyl)thio]butylamine was obtained as an oil.

NMR (δ ppm in CDCl₃): 1.37 (2H, s), 1.52-1.67 (2H, m), 1.82 (2H, m),2.74 (2H, t, J=7 Hz ), 3.24 (2H, t, J=7 Hz ), 7.21 (1H, d, J=3 Hz), 7.67(1H, d, J=3 Hz).

Reference Example 58

According to the same manner as that described in Reference Example 27,4-[(1-methyl-1,2,3,4-tetrazol-5-yl) thio]butylamine was obtained as anoil.

NMR (δ ppm in CDCl₃): 1.61 (2H, m), 1.69 (2H, s), 1.88 (2H, m), 2.77(2H, t, J=7 Hz ), 3.37 (2H, t, J=7 Hz ), 3.92 (3H, s).

EXAMPLE 1

4-(Phenylthio)butylamine (7.90 g), ethyl orthoformate (12.9 g) anddiethyl phosphite (24.1 g) were stirred overnight at 150° C. Thereaction mixture was concentrated under reduced pressure and the residuewas subjected to silica gel column chromatography. Tetraethyl4-(phenylthio)butylaminomethylene bisphosphonate (11.1 g, 54%) wasobtained as a colorless oil from fraction eluted withchloroform-methanol (50:1, v/v).

NMR (δ in CDCl₃): 1.34 (12H, t, J=7 Hz), 1.50-1.70 (4H, m), 2.85 (2H, t,J=7 Hz), 2.93 (2H, t, J=7 Hz), 3.23 (1H, t, J=22 Hz), 4.12-4.30 (8H, m),7.10-7.40 (5H, m).

EXAMPLE 2

According to the same manner as that in Example 1, tetraethyl4-(4-chlorophenylthio)butylaminomethylenebisphosphonate was obtained asa colorless oil starting from 4-(4-chlorophenylthio)butylamine (40%).

NMR δ (CDCl₃): 1.34 (12H, t, J=7 Hz), 1.55-1.70 (4H, m), 2.86 (2H, t,J=7 Hz), 2.90 (2H, t, J=7 Hz), 3.23 (1H, t, J=22 Hz), 4.12-4.29 (8H, m),7.24 (4H, s).

EXAMPLE 3

According to the same manner as that in Example 1, tetraethyl3-(phenylthio)propylaminomethylenebisphosphonate was obtained as acolorless oil starting from 3-(phenylthio)propylamine (33%).

NMR δ (CDCl₃): 1.35 (12H, t, J=7 Hz), 1.65-1.80 (2H, m), 2.95 (2H, d,J=7 Hz), 3.01 (2H, d, J=7 Hz), 3.27 (1H, t, J=22 Hz), 4.10-4.30 (8H, m),7.10-7.40 (5H, m).

EXAMPLE 4

According to the same manner as that in Example 1, tetraethyl2-(phenylthio)ethylaminomethylenebisphosphonate was obtained as acolorless oil starting from 3-(phenylthio)ethylamine (28%).

NMR δ (CDCl₃): 1.33 (12H, t, J=7 Hz), 3.5 (4H, s), 3.27 (1H, t, J=22Hz), 4.12-4.29 (8H, m), 7.14-7.40 (5H, m).

EXAMPLE 5

Tetraethyl 4-(phenylthio)butylaminomethylenebisphosphonate (11.1 g) wasdissolved in concentrated hydrochloric acid (150 ml) and the mixture wasrefluxed for 2.5 hours. After the reaction mixture was concentratedunder reduced pressure, water was poured and separated crystals werecollected by filtration to obtain4-(phenylthio)butylaminomethylenebisphosphonic acid (4.94 g) as a whitepowder. Sodium methylate (28% methanol solution, 13.3 g) was added tothe suspension of this powder (4.08 g) in methanol (50 ml), and theresultant was stirred at room temperature for one hour and thenconcentrated under reduced pressure. Methanol was added to the residue,the separated white crystals were collected by filtration andrecrystallized from water-methanol to obtain tetrasodium4-(phenylthio)butylaminomethylenebisphosphonate as a white powder (3.90g, 43% ).

Melting Point: >300° C.

Elemental Analysis for C₁₁ H₁₅ NO₆ SP₂ Na₄.H₂ O, Calcd.: C, 28.65; H,3.72; N, 3.04 Found: C, 28.50; H, 3.69; N, 2.95

NMR δ (D₂ O): 1.67-1.86 (4H, m), 2.91 (1H, t, J=17 Hz), 3.06 (2H, t, J=7Hz), 3.24 (2H, t, J=7 Hz), 7.25-7.50 (5H,m).

EXAMPLE 6

According to the same manner as that in Example 5, tetrasodium4-(4-chlorophenylthio)butylaminomethylenebisphosphonate (68%,recrystallization solvent: water-MeOH) was obtained as a white powderstarting from tetraethyl4-(4-chlorophenylthio)butylaminomethylenebisphosphonate.

Elemental Analysis for C₁₁ H₁₄ ClNO₆ SP₂ Na₄.1/2H₂ O, Calcd.: C, 27.15;H, 3.11; N, 2.88 Found: C, 27.28; H, 3.40; N, 2.94

NMR δ (D₂ O): 1.65-1.90 (4H, m), 2.90 (1H, t, J=17 Hz), 3.04 (2H, t, J=7Hz), 3.23 (2H, t, J=7 Hz), 7.39 (4H, s).

EXAMPLE 7

According to the same manner as that in Example 5, tetrasodium3-(phenylthio) propylaminomethylenebisphosphonate (45%,recrystallization solvent: water-MeOH) was obtained starting fromtetraethyl 3-(phenylthio) propylaminomethylenebisphosphonate.

Elemental Analysis for C₁₀ H₁₃ NO₆ SP₂ Na₄.H₂ O, Calcd.: C, 26.86; H,3.38; N, 3.13 Found: C, 27.12; H, 3.48; N, 2.97

NMR δ (D₂ O): 2.00 (2H, quintet, J=7 Hz), 2.89 (1H, t, J=17 Hz), 3.10(2H, d, J=7 Hz ), 3.32 (2H, d, J=7 Hz), 7.25-7.55 (5H, m).

Example 8

According to the same manner as that in Example 5, tetrasodium2-(phenylthio)ethylaminomethylenebisphosphonate as a white powder wasobtained starting from tetraethyl2-(phenylthio)ethylaminomethylenebisphosphonate (43%, recrystallizationsolvent: water-methanol).

Melting Point: >300° C.

Elemental Analysis for C₉ H₁₁ NO₆ SP₂ Na₄.1/2H₂ O, Calcd.: C, 25.48; H,2.85; N, 3.30 Found: C, 25.37; H, 2.90; N, 3.19

NMR δ (D₂ O): 2.75 (1H, t, J=17 Hz), 3.24 (4H, s), 7.27-7.57 (5H, m).

Example 9

According to the same manner as that in Example 1, tetraethyl4-(2-pyridylthio)butylaminomethylenebisphosphonate was obtained as acolorless oil starting from 4-(2-pyridylthio)butylamine (yield: 54%).

NMR δ (CDCl₃): 1.34 (12H, t, J=7 Hz), 1.60-1.90 (4H, m), 2.88(2H, t, J=7Hz), 3.18 (2H, t, J=7 Hz), 3.26 (1H, t, J=22 Hz), 4.10-4.30 (8H, m),6.96 (1H, ddd, J=7, 5, 1 Hz), 7.16 (1H, d, J=8 Hz), 7.47 (1H, ddd, J=8,7, 2 Hz), 8.42 (1H, ddd, J=5, 2, 1 Hz).

Example 10

A solution of tetraethyl4-(2-pyridylthio)butylaminomethylenebisphosphonate (2.50 g) inhydrochloric acid (40 ml) was stirred for 3 hours under reflux. Thereaction mixture was concentrated under reduced pressure. Acetone wasadded to the residue and the separated white precipitate was collectedby filtration and recrystallized from water-methanol to obtain4-(2-pyridylthio)butylaminomethylenebisphosphonic acid hydrochloride(1.46 g, 70%) as a white powder.

Melting Point: 176-178° C.

Elemental Analysis for C₁₀ H₁₈ N₂ O₆ SP₂.HCl, Calcd.: C, 30.58; H, 4.88;N, 7.13 Found: C, 30.59; H, 5.10; N, 7.02

NMR δ (D₂ O): 1.80-2.00 (4H, m), 3.35-3.47 (4H, m), 3.53 (1H, t, J=17Hz), 7.70 (1H, dd, J=7, 6 Hz), 7.95 (1H, J=8 Hz), 8.36 (1H, dd, J=8, 7Hz), 8.53 (1H, d, J=6 Hz).

EXAMPLE 11-31

According to the same manner as that in Example 1, the compounds shownin Tables 12-15 were obtained as oil.

                                      TABLE 12                                    __________________________________________________________________________     ##STR78##                                                                     Ex. No.                                                                            ##STR79##                                                                            n  (°C.)Reaction temp.                                                            (hr.)Reaction time                                                                    Yield (%)                                                                           NMR (δppm in                      __________________________________________________________________________                                         CDCl.sub.3)                              11                                                                                  ##STR80##                                                                           5  150     5       44    1.35(12H, t, J=7Hz), 1.42-1.75(6H,                                            m), 2.83(2H, t, J=7Hz), 2.91(2H, t,                                           J=7Hz), 3.24(1H, t, J=22Hz), 4.10-                                            4.32(8H, m), 7.12-7.36(5H, m)            12                                                                                  ##STR81##                                                                           6  150     4       40    1.34(12H, t, J=7Hz), 1.40-1.80(8H,                                            m), 2.91(4H, t, J=7Hz), 3.24(1H, t,                                           J=22Hz), 4.10-4.30(8H, m), 7.10-                                              7.37(5H, m)                              13                                                                                  ##STR82##                                                                           2  150     2       28    1.33(12H, t, J=7Hz), 3.15(2H, t, J=                                           7Hz), 3.31(2H, t, J=7Hz), 3.40(1H,                                            t, J=22Hz), 4.11-4.31(8H, m), 6.97                                            (1H, ddd, J=7, 5, 1Hz), 7.17(1H,                                              ddd, J=8, 1, 1Hz), 7.46(1H, ddd,                                              J=8, 7, 2Hz), 8.40(1H, ddd, J=5, 2,                                           1Hz)                                     14                                                                                  ##STR83##                                                                           3  150     2       47    1.34(12H, t, J=7Hz), 1.88(2H,                                                 quintet, J=7Hz), 3.00(2H, t, J=7Hz)                                           3.25(2H, t, J=7Hz), 3.31(1H, t, J=                                            22Hz), 4.11-4.31(8H, m), 6.97(1H,                                             ddd, J=7, 5, 1Hz), 7.16(1H, ddd, J=                                           8, 1, 1Hz), 7.47(1H, ddd, J=8, 7,                                             2Hz) 8.43(1H, ddd, J=5, 2, 1Hz)          15                                                                                  ##STR84##                                                                           5  150     3       45    1.35(12H, t, J=7Hz), 1.45-1.80(6H,                                            m), 2.84(2H, t, J=7Hz), 3.16(2H, t,                                           J= 7Hz), 3.24(1H, t, J=22Hz),                                                 4.14-4.30 (8H, m), 6.96(1H, ddd,                                              J=7, 5, 1Hz), 7.15(1H, ddd, J=8, 1,                                           1Hz), 7.46 (1H, ddd, J=8, 7, 2Hz),                                            8.41(1H, ddd, J=5, 2,                    __________________________________________________________________________                                         1Hz)                                 

                                      TABLE 13                                    __________________________________________________________________________     ##STR85##                                                                     Ex. No.                                                                            ##STR86##                                                                            n  (°C.)Reaction temp.                                                            (hr.)Reaction time                                                                    Yield (%)                                                                           NMR (δppm in                      __________________________________________________________________________                                         CDCl.sub.3)                              16                                                                                  ##STR87##                                                                           6  150     3       27    1.35(12H, t, J=7Hz), 1.30-1.80(8H,                                            m), 2.83(2H, t, J=7Hz), 3.16(2H, t,                                           J=7Hz), 3.25(1H, t, J=22Hz), 4.13-                                            4.32(8H, m), 6.97(1H, ddd, J=7, 5, 1                                          z), 7.17(1H, ddd, J=8, 1, 1Hz), 7.47                                          1H, ddd, J=8, 7, 2Hz), 8.42(1H, ddd,                                          =5, 2, 1Hz)                              17                                                                                  ##STR88##                                                                           2  145     2       16    1.34(12H, t, J=7Hz), 1.95(1H, s),                                             3.1-3.3(4H, m), 3.29(1H, t, J=22Hz),                                          .1-4.3(8H, m), 7.13(2H, d, J=6Hz),                                            8.39(2H, d, J=6Hz)                       18                                                                                  ##STR89##                                                                           3  145     2       21    1.35(12H, t, J=7Hz), 1.7-2.0(3H, m)                                           2.9-3.2(4H, m), 3.25(1H, t, J=22Hz),                                          .1-4.3(8H, m), 7.12(2H, double d,                                             J=6 and 2Hz), 8.38(2H, double d, J=6                                          and 2Hz)                                 19                                                                                  ##STR90##                                                                           4  150     2       10    1.34(12H, t, J=7Hz), 1.56-1.86(4H,                                            m), 2.90(2H, t, J=7Hz), 3.00(2H, t,                                           J=7Hz), 3.24(1H, t, J=22Hz), 4.08-                                            4.31(8H, m), 7.11(2H, dd, J=6, 2Hz),                                          .39(2H, dd, J=6, 2Hz)                    20                                                                                  ##STR91##                                                                           5  140     3       25    1.35(12H, t, J=7Hz), 1.4-1.6(2H, m)                                           1.6-1.8(2H, m), 1.9-2.2(3H, m),                                               2.85(2H, t, J=7Hz), 3.00(2H, t, J=                                            7Hz), 3.24(1H, t, J=22Hz), 4.1-4.3                                            (8H, m), 7.10(2H, d, J=6Hz),                                                  8.38(2H, d, J=6Hz)                       __________________________________________________________________________

                                      TABLE 14                                    __________________________________________________________________________     ##STR92##                                                                     Ex. No.                                                                            ##STR93##                                                                               n                                                                               (°C.)Reaction temp.                                                            (hr.)Reaction time                                                                   Yield (%)                                                                           NMR (δppm in                     __________________________________________________________________________                                          CDCl.sub.3)                             21                                                                                  ##STR94##                                                                              2 140     3      30    1.34(12H, t, J=7Hz), 1.92(1H, s),                                             3.12-3.21(4H, m), 3.31(1H, t, J=22                                            Hz), 3.62(3H, s), 4.13-4.29(8H, m)                                            6.91(1H, d, J=1Hz), 7.03(1H, d,                                               J=1Hz)                                  22                                                                                  ##STR95##                                                                              4 140     3      11    1.35(12H, t, J=7Hz), 1.63-1.73(4H,                                            m), 1.88(1H, s), 2.85(2H, t,                                                  J=7Hz), 3.10(2H, t, J=7Hz),                                                   3.23(1H, t, J=22 Hz), 3.62(3H, s),                                            4.13-4.28(8H, m), 6.93(1H, d,                                                 J=1Hz), 7.07(1H, d, J=1Hz)              23                                                                                  ##STR96##                                                                              2 150     3      38    1.35(12H, t, J=7Hz), 1.79(1H, s),                                             2.72(3H, s), 3.27(2H, t, J=6Hz),                                              3.32 (1H, t, J=21Hz), 3.44(2H, t,                                             J=6Hz), 4.14-4.31(8H, m)                24                                                                                  ##STR97##                                                                              4 140     3      40    1.35(12H, t, J=7Hz), 1.59-1.67(2H,                                            m), 1.67(1H, s), 1.82-1.90(2H, m),                                            2.72(3H, s), 2.88(2H, t, J=7Hz),                                              3.24 (1H, t, J=22Hz), 3.32(2H, t,                                             J=7Hz), 4.14-4.29(8H, m)                25                                                                                  ##STR98##                                                                              4 140     3      43    1.34(12H, t, J=8Hz), 1.64-1.81(5H,                                            m), 2.90(2H, t, J=7Hz), 3.15(2H, t,                                           J= 7Hz), 3.26(1H, t, J=22Hz),                                                 4.13-4.28 (8H, m), 6.95(1H, t,                                                J=5Hz), 8.52(2H, d J=5Hz)               26                                                                                  ##STR99##                                                                              3 140     3      24    1.35(12H, t, J=7Hz), 1.73(1H, s),                                             1.91(2H, m), 3.00(2H, t, J=7Hz),                                              3.23 (2H, t, J=7Hz), 3.31(IH, t,                                              J=22Hz), 4.14-4.30(8H, m), 6.95(1H,                                           t, J=5Hz) 8.5(2H, d,                    __________________________________________________________________________                                          J=5Hz)                              

                                      TABLE 15                                    __________________________________________________________________________     ##STR100##                                                                    Ex. No.                                                                            ##STR101##                                                                              n                                                                               (°C.)Reaction temp.                                                            (hr.)Reaction time                                                                   Yield (%)                                                                           NMR (δppm in                     __________________________________________________________________________                                          CDCl.sub.3)                             27                                                                                  ##STR102##                                                                             2 140     3      27    1.34(12H, t, J=7Hz), 1.69(1H, s),                                             3.17-3.32(4H, m), 3.40(1H, t, J=22                                            Hz), 4.14-4.28(8H, m), 6.97(1H, t,                                            J=5Hz), 8.51(2H, d, J=5Hz)              28                                                                                  ##STR103##                                                                             4 150     3      24     1.35(12H, t, J=7Hz), 1.57-1.77(5H,                                           ), 2.86(2H, t, J=7Hz), 3.11(2H, t,                                            J= 7Hz), 3.24(1H, t, J=22Hz),                                                 3.38(2H, t, J=8Hz), 4.21(2H, t,                                               J=8Hz), 4.14- 4.30(8H, m)               29                                                                                  ##STR104##                                                                             4 150     5      18    1.34(12H, t, J=7Hz), 1.5- 2.00(4H,                                            m), 2.88(2H, t, J=7Hz), 3.20(1H, t,                                           =22Hz), 3.22(2H, t, J=7Hz), 4.05-                                             4.30(8H, m), 6.93-7.10(1H, m), 7.16                                           1H, dd, J=8 and 1Hz), 7.40-7.46                                               (1H, m), 8.39-8.45(1H, m)               30                                                                                  ##STR105##                                                                             4 140     3      28    1.34(12H, t, J=7Hz), 1.5-2.0(5H, m)                                           .85(2H, t, J=7Hz), 3.18(2H, t, J=                                             7Hz), 3.23(1H, t, J=22Hz), 4.1-                                               4.3(8H, m), 6.93(1H, t, J=6Hz),                                               7.25- 7.35(1H, m), 7.77(1H, d,                                                J=6Hz), 8.11(1H, d, J=6Hz)              31                                                                                  ##STR106##                                                                             5 150     3      43    1.35(12H, t, J=7Hz), 1.48-1.55(3H,                                            m), 1.67-1.79(4H, m), 2.85(2H, t,                                             J=6Hz), 3.14(2H, t, J=7Hz), 3.25                                              (1H, t, J=22Hz), 4.14-4.30(8H, m),                                            6.96(1H, t, J=5Hz), 8.51(2H, d,                                               J=5Hz)                                  __________________________________________________________________________

EXAMPLE 32

m-Chloroperbenzoic acid (487 mg) was added in

Small portions to a solution of tetraethyl4-(phenylthio)butylaminomethylenebisphosphonate (1.20 g) indichloromethane (10 ml) with ice-cooling, and then the mixture wasstirred at 0° C. for 2 hours. The reaction mixture was washedsuccessively with aqueous sodium bisulfite, saturated aqueous sodiumbicarbonate, and water and then dried (MgSO₄), and concentrated. Theresidue was chromatographed on silica gel. Tetraethyl4-(phenylsulfinyl)butylaminomethylenebisphosphonate was obtained as acolorless oil (1.05 g, 85%) from the fraction eluted withchloroformmethanol (50:1, v/v).

NMR (δ ppm in CDCl₃): 1.34 (12H, t, J=7 Hz ), 1.45-2.00 (4H, m), 2.82(2H, t, J=7 Hz ), 2.85 (2H, t, J=7 Hz ), 3.21 (1H, t, J=22 Hz),4.08-4.32 (8H, m), 7.48-7.67 (5H, m).

EXAMPLES 33-35

According to the same manner as that in Example 32, the compounds shownin Table 16 were obtained.

                                      TABLE 16                                    __________________________________________________________________________     ##STR107##                                                                    Ex. No.                                                                            ##STR108##                                                                           k                                                                               Yield (%)                                                                           NMR (δppm in CDCl.sub.3)                           __________________________________________________________________________    33                                                                                  ##STR109##                                                                          2 29    1.34(12H, t, J=7Hz), 1.50-1.95(4H, m), 2.84(2H, t,                            J=7Hz), 3.13(2H, t, J=7Hz), 3.19(1H, t, J=22Hz),                              4.08-4.30(8H, m), 7.52-7.74(3H, m), 7.86-7.96(2H, m)      34                                                                                  ##STR110##                                                                          1 83    1.34(12H, t, J=7Hz), 1.50-2.00(4H, m), 2.85(2H, t,                            J=7Hz), 3.11(2H, t, J=7Hz), 3.22(1H, t, J=22Hz),                              4.06-4.29(8H, m), 7.38(1H, ddd, J=7, 5, 1Hz),                                 7.87-7.99  (2H, m), 8.62(1H, ddd, J=5, 2, 1Hz)            35                                                                                  ##STR111##                                                                          2 34    1.33(12H, t, J=7Hz), 1.50-2.00(4H, m), 2.85(2H, t,                            J=7Hz), 3.21(1H, t, J=22Hz), 3.43(2H, t, J=7Hz),                              4.00-4.30(8H, m), 7.37(1H, ddd, J=7, 5, 1Hz),                                 7.86-8.04(2H, m), 8.61(1H, ddd, J=5, 2,                   __________________________________________________________________________                        1Hz)                                                  

EXAMPLES 36-39

According to the same manner as that in Example 5, the compounds shownin Table 17 were obtained.

                                      TABLE 17                                    __________________________________________________________________________     ##STR112##                                                                    Ex. No.                                                                            ##STR113##                                                                           m                                                                               n                                                                               Yield (%)                                                                           m.p. (°C.)                                                                   Recrystn. solvent                                                                      Molecular formula                       __________________________________________________________________________    36                                                                                  ##STR114##                                                                          1 4 49    >300  water-MeOH                                                                             C.sub.11 H.sub.15 NO.sub.7 P.sub.2                                            SNa.sub.4.2H.sub.2 O                     37                                                                                  ##STR115##                                                                          2 4 73    >300  water-MeOH                                                                             C.sub.11 H.sub.15 NO.sub.8 P.sub.2                                            SNa.sub.4.H.sub.2 O                      38                                                                                  ##STR116##                                                                          0 5 48    >300  water-MeOH                                                                             C.sub.12 H.sub.17 NO.sub. 6 P.sub.2                                           SNa.sub.4                                39                                                                                  ##STR117##                                                                          0 6 54    >300  water-MeOH                                                                             C.sub.13 H.sub.19 NO.sub.6 P.sub.2                                            SNa.sub.4                                __________________________________________________________________________

EXAMPLE 40

Bromotrimethylsilane (1.80 g) was added dropwise to a solution oftetraethyl 4-[(2-pyridine 1-oxide)thio]butylaminomethylenebisphosphonate(1.14 g) in acetonitrile (20 ml) and the resultant was stirred overnightat room temperature. Water was added to the reaction mixture, and theresultant was further stirred at room temperature for one hour, and thenconcentrated under reduced pressure. The residue was dissolved inmethanol (10 ml), to which was added sodium methoxide (28% methanolsolution, 2.72 g), and the resultant was stirred at room temperature forone hour and concentrated under reduced pressure. Acetone was added tothe residue and the separated white precipitate was collected byfiltration, which was recrystallized from water-methanol to obtaintrisodium 4-[(2-pyridine 1-oxide)thio]butylaminomethylenebisphosphonateas a white powder (220 mg, 20%).

Melting Point: >300° C.

Elemental Analysis for C₁₀ H₁₅ N₂ O₇ P₂ SNa₃.1.5H₂ O, Calcd.: C, 25.82;H, 3.90; N, 6.02 Found: C, 26.16; H, 3.80; N, 5.88

EXAMPLE 41

Bromotrimethylsilane (13.8 g) was added to a solution of tetraethyl4-(2-pyrimidylthio)butylaminomethylenebisphosphonate (7.0 g) inacetonitrile (70 ml) and the resultant was stirred at room temperaturefor 15 hours. After water (5 ml) was added, the reaction mixture wasconcentrated under reduced pressure. A solution of sodium methoxide inmethanol (28%, 18.0 g) and ether (100 ml) were added to the residue, andthe separated crystals were collected by filtration and recrystallizedfrom water-ethanol to obtain tetrasodium4-(2-pyrimidylthio)butylaminomethylenebisphosphonate monohydrate (2.1 g,30%) as colorless prisms.

Melting Point: >300° C.

Elemental Analysis for C₉ H₁₃ N₃ O₆ P₂ SNa₄.H₂ O, Calcd.: C, 23.34; H,3.26; N, 9.07 Found: C, 23.69; H, 3.57; N, 8.95

EXAMPLES 42-50

According to the same manner as that in Example 41, the compounds shownin Table 18 were obtained.

                                      TABLE 18                                    __________________________________________________________________________     ##STR118##                                                                    Ex. No.                                                                            ##STR119##                                                                              n                                                                               Yield (%)                                                                           m.p. (°C.)                                                                   Recrystn. solvent                                                                      Molecular formula                      __________________________________________________________________________    42                                                                                  ##STR120##                                                                             4 26    >300  water-MeOH                                                                             C.sub.9 H.sub.15 N.sub.3 O.sub.6                                              P.sub.2 SNa.sub.4.H.sub.2 O             43                                                                                  ##STR121##                                                                             2 29    >300  water-MeOH                                                                             C.sub.7 H.sub.11 N.sub.3 O.sub.6                                              P.sub.2 SNa.sub.4.7H.sub.2 O            44                                                                                  ##STR122##                                                                             4 41    >300  MeOH-ether                                                                             C.sub.8 H.sub.13 N.sub.3 O.sub.6                                              P.sub.2 S.sub.2 Na.sub.4.4H.sub.2                                             O                                       45                                                                                  ##STR123##                                                                             2 43    >300  water-MeOH                                                                             C.sub.6 H.sub.9 N.sub.3 O.sub.6                                               P.sub.2 S.sub.2 Na.sub.4.6H.sub.2                                             O                                       47                                                                                  ##STR124##                                                                             5 40    >300  water-MeOH                                                                             C.sub.10 H.sub.15 N.sub.3 O.sub.6                                             P.sub.2 SNa.sub.4.H.sub.2 O             48                                                                                  ##STR125##                                                                             3 23    >300  water-MeOH                                                                             C.sub.8 H.sub.11 N.sub.3 O.sub.6                                              P.sub.2 SNa.sub.4.2H.sub.2 O            49                                                                                  ##STR126##                                                                             2 57    >300  water-MeOH                                                                             C.sub.7 H.sub.9 N.sub.3 O.sub.6                                               P.sub.2 SNa.sub.4.1.5H.sub.2 O          50                                                                                  ##STR127##                                                                             4 32    >300  water-MeOH                                                                             C.sub.8 H.sub.14 N.sub.2 O.sub.6                                              P.sub.2 S.sub.2 Na.sub.4.0.5H.sub.2                                           O                                       __________________________________________________________________________

EXAMPLE 51

A mixture of tetraethyl3-(4-pyridylthio)propylaminomethylenebisphosphonate (7.4 g) andconcentrated hydrochloric acid (50 ml) was heated under reflux for 2hours, and then concentrated under reduced pressure. The residualcrystals were collected by filtration to obtain3-(4-pyridylthio)propylaminomethylenebisphosphonic acid hemihydrate (4.3g, 77%), and recrystallized from water.

Melting Point: 280°-281° C.

Elemental Analysis for C₉ H₁₆ N₂ O₆ P₂ S.1/2H₂ O, Calcd.: C, 30.77; H,4.87; N, 7.98 Found: C, 30.60; H, 5.09; N, 7.92

EXAMPLES 52-60

According to the same manner as that in Example 51, the compounds shownin Table 19 were obtained.

                                      TABLE 19                                    __________________________________________________________________________     ##STR128##                                                                    Ex. No.                                                                            ##STR129##                                                                             m                                                                               n                                                                               Yield (%)                                                                           m.p. (°C.)                                                                   Recrystn. solvent                                                                      Molecular formula                     __________________________________________________________________________    52                                                                                  ##STR130##                                                                            0 2 68    252-253                                                                             water-MeOH                                                                             C.sub.8 H.sub.14 N.sub.2 O.sub.6                                              P.sub.2 S.0.5H.sub.2 O                 53                                                                                  ##STR131##                                                                            0 3 68    242-244                                                                             water-MeOH                                                                             C.sub.9 H.sub.16 N.sub.2 O.sub.6                                              P.sub.2 S.0.5H.sub.2 O                 54                                                                                  ##STR132##                                                                            0 5 66    208-210                                                                             water-MeOH                                                                             C.sub.11 H.sub.20 N.sub.2 O.sub.6                                             P.sub.2 S.H.sub.2 O                    55                                                                                  ##STR133##                                                                            0 6 61    213-214                                                                             water-MeOH                                                                             C.sub.12 H.sub.22 N.sub.2 O.sub.6                                             P.sub.2 S                              56                                                                                  ##STR134##                                                                            0 2 93    288-289                                                                             water    C.sub.8 H.sub.14 N.sub.2 O.sub.6                                              P.sub.2 S.0.5H.sub.2 O                 58                                                                                  ##STR135##                                                                            0 5 95    279-280                                                                             water    C.sub.11 H.sub.20 N.sub.2 O.sub.6                                             P.sub.2 S.0.5H.sub.2 O                 59                                                                                  ##STR136##                                                                            0 4 39    222-223                                                                             water    C.sub.11 H.sub.18 N.sub.4 O.sub.6                                             P.sub.2 S.0.5H.sub.2 O                 60                                                                                  ##STR137##                                                                            1 4 47    232-235                                                                             water-MeOH                                                                             C.sub.10 H.sub.18 N.sub.2 O.sub.7                                             P.sub.2 S.1.5H.sub.2 O                 __________________________________________________________________________

EXAMPLE 61

5-(2-Pyridythio)pentylaminomethylenebisphosphonic acid monohydrate (0.75g) was suspended in methanol (20 ml) and a solution of sodium methoxidein methanol (28%, 1.0 g) was added thereto. The resultant was stirred atroom temperature for 5 hours. The reaction mixture was concentratedunder reduced pressure and the residual solid was recrystallized fromwater-methanol to obtain disodium5-(2-Pyridylthio)pentylaminomethylenebisphosphonate 1.5 hydrate (0.591g, 69%).

Melting Point: >300° C.

Elemental Analysis for C₁₁ H₁₈ N₂ O₆ P₂ SNa₂.1.5H₂ O, Calcd.: C, 29.94;H, 4.80; N, 6.35 Found: C, 30.05; H, 4.58; N, 6.36

EXAMPLES 62-65

According to the same manner as that in Example 61, the compounds shownin Table 20 were obtained.

                                      TABLE 20                                    __________________________________________________________________________     ##STR138##                                                                    Ex. No.                                                                            ##STR139##                                                                           n                                                                               Yield (%)                                                                           m.p. (°C.)                                                                   Recrystn. solvent                                                                      Molecular formula                         __________________________________________________________________________    62                                                                                  ##STR140##                                                                          2 77    >300  water-MeOH                                                                             C.sub.8 H.sub.12 N.sub.2 O.sub.6                                              P.sub.2 SNa.1.5H.sub.2 O                   63                                                                                  ##STR141##                                                                          3 73    >300  water-MeOH                                                                             C.sub.9 H.sub.14 N.sub.2 O.sub.6                                              P.sub.2 SNa.sub.2.1.5H.sub.2 O             65                                                                                  ##STR142##                                                                          6 70    >300  water-MeOH                                                                             C.sub.12 H.sub.20 N.sub.2 O.sub. 6                                            P.sub.2 SNa.sub.2.0.5H.sub.2 O             __________________________________________________________________________

EXAMPLE 66

According to the same manner as that in Example 1, tetraethyl4-[(4-methoxyphenyl)thio]butylaminomethylenebisphosphonate was obtainedas an oil.

NMR (δ ppm in CDCl₃): 1.35 (12H, t, J=7 Hz), 1.58-1.90 (4H, m), 2.91(2H, t, J=7 Hz), 3.27 (12H, t, J=22 Hz), 3.77 (3H, s), 3.91 (2H, t, J=7Hz), 4.12-4.30 (8H, m), 6.82 (4H, s).

EXAMPLE 67

According to the same manner as that in Example 1, tetraethyl4-[(2-thiazolyl)thio]butylaminomethylenebisphosphonate was obtained asan oil.

NMR (δ ppm in CDCl₃): 1.34 (12H, t, J=7 Hz), 1.58-1.69 (3H, m),1.76-1.87 (2H, m), 2.88 (2H, t, J=7 Hz), 3.23 (2H, t, J=7 Hz), 3.24 (1H,t, J=22 Hz), 4.13-4.29 (8H, m), 7.21 (1H, d, J=3 Hz), 7.66 (1H, d, J=3Hz).

Example 68

According to the same manner as that in Example 1, tetraethyl4-[(1-methyl-1 , 2, 3,4-tetrazol-5-yl)thio]butylaminomethylenebisphosphonate was obtained asan oil

NMR (δ ppm in CDCl₃): 1.35 (12H, t, J=7 Hz), 1.59-1.93 (5H, m), 2.89(2H, t, J=7 Hz), 3.24 (1H, t, j=22 Hz ), 3.38 (2H, t, J=7 Hz), 3.92 (3H,s), 4.14-4.31 (8H, m).

EXAMPLE 69

According to the same manner as that in Example 55,4-[(4-methoxyphenyl)thio]butylaminomethylenebisphosphonic acid wasobtained, m.p. 188°-189° C.

Elemental analysis for C₁₂ H₂₁ NO₇ P₂ S.H₂ O, Calcd.: C, 37.22; H, 5.99;N, 3.62 Found: C, 36.99; H, 6.19; N, 3.74

Example 70

According to the same manner as that in Example 61, disodium4-[(4-methoxyphenyl)thio]butylaminometnylenebisphosphonate was obtainedand recrystallized from water-methanol, m.p. >300° C.

Elemental analysis for C₁₂ H₁₉ NO₇ P₂ SNa₂.2H₂ O, Calcd.: C, 32.08; H,5.16; N, 3.12 Found: C, 32.05; H, 5.06; N, 3.26

EXAMPLE 71

To a solution of tetraethyl4-[(2-thiazolyl)thio]butylaminomethylenebisphosphonate (6.34 g) inacetonitrile (100 ml ) was added bromotrimethylsilane (12.27 g) and themixture was stirred at room temperature for 15 hours. Water (3.3 ml) wasadded to the reaction mixture and the mixture was concentrated underreduced pressure. The residue was suspended in methanol (50 ml) andsodium methylate

(28% methanol solution, 15.5 ml ) was added thereto. The mixture wastreated with ether (150 ml), and the separated solid was filtered offand recrystallized from water-methanol to obtain disodium4-[(2-thiazolyl]thio]butylaminomethylenebisphosphonate (3.36 g),m.p. >300° C.

Elemental analysis for C₈ H₁₄ N₂ O₆ P₂ S₂ Na₂.2.5H₂ O, Calcd.: C, 21.29;H, 4.24; N, 6.21 Found: C, 21.11; H, 4.42; N, 6.08

EXAMPLE 72

According to the same manner as that in Example 40, tetrasodium4-[(1-methyl-1, 2, 3,4-tetrazol-5-yl)thio]butylaminomethylenebisphosphonate was obtained andrecrystallized from water-methanol, m.p. >300° C.

Elemental analysis for C₇ H₁₅ N₂ O₇ P₂ Na₄, Calcd.: C, 18.00; H, 3.24;N, 14.99 Found: C, 18.25; H, 3.62; N, 14.70

Example 73

According to the same manner as that in Example 10,4-[(2-pyridyl)sulfinyl]butylaminomethylenebisphosphonic acid wasobtained and recrystallized from water-ethanol, m.p. 235-240° C.

Elemental analysis for C₁₀ H₁₈ N₂ O₇ P₂ S.H₂ O, Calcd.: C, 30.77; H,5.17; N, 6.67 Found: C, 30.87; H, 5.28; N, 6.79

EXAMPLE 74

To a solution of tetraethyl4-(phenylthio)butylaminomethylenebisphosphonate (2.5 g) in ethanol (20ml) was added a solution of sodium hydroxide (450 mg) in ethanol (20ml). The mixture was heated under reflux for 4 hours and concentratedunder reduced pressure. The residue was dissolved in water and thesolution was subjected to column chromatography on Amberlite CG-50 (H⁺form) and eluted with water to obtain diethyl4-(phenylthio)butylaminomethylenebisphosphonate monosodium salt (810mg). The resultant was recrystallized from methanol-hexane, m.p.143°-145° C.

NMR (δ ppm in D₂ O): 1.27 (6H, t, J=7 Hz), 1.65-1.95 (4H, m), 3.05 (2H,t, J=7 Hz), 3.3-3.4 (2H, m), 3.46 (1H, t, J=18.5 Hz), 3.9-4.1 (4H, m),7.25-7.5 (5H, m).

Elemental analysis for C₁₅ H₂₆ NO₆ P₂ SNa.1/2H₂ O, Calcd.: C, 40.73; H,6.15; N, 3.17 Found: C, 40.70; H, 6.18; N, 3.25

EXAMPLE 75

To a solution of tetraethyl4-(phenylthio)butylaminomethylenebisphosphonate (1.0 g) in acetonitrile(10 ml ) was added bromotrimethylsilane (0.98 g) and the mixture wasstirred at room temperature for 2 days. Water was added to the reactionmixture and concentrated under reduced pressure. The residual solid wasfiltered off and recrystallized from H₂ O--CH₃ OH to obtain ethyl4-(phenylthio)butylaminomethylene-bisphosphonate (3.36 g), m.p.189°-190° C.

NMR (δ ppm in d₆ -DMSO ): 1.17 (3H, t, J=7 Hz), 1.5-1.9 (4H, m), 2.96(2H, t, J=7 Hz), 3.16 (2H, broad t, J=7 Hz), 3.36 (1H, t, J=18 Hz),3.8-4.0 (2H, m), 7.1-7.3 (5H, m), 7.48 (3H, broad s ).

Elemental analysis for C₁₃ H₂₃ NO₆ P₂ S.1/2H₂ O, Calcd.: C, 39.80; H,6.17; N, 3.57 Found: C, 39.93; H, 6.00; N, 3.66

Example 76

Ethyl 4-(phenylthio)butylaminomethylenebisphosphonate hemihydrate (0.2g) was dissolved in 1N NaOH (0.5 ml) and pH of the solution was adjustedto 7 with 1N NaOH. The solution was subjected to column chromatographyon a Amberlite CG-50 (H⁺ form) column and eluted with water to obtainethyl 4-(phenylthio)butylaminomethylenebisphosphate monosodium salt (120mg). The resultant was recrystallized from water-ethanol, m.p. 167°-169°C.

NMR (δ ppm in D₂ O): 1.26 (3H, t, J=7 Hz ) 1.65-2.0 (4H, m), 3.05 (2H,t, J=7 Hz), 3.3-3.45 (2H, m), 3.47 (1H, t, J=18 Hz), 3.9-4.1 (4H, m),7.25-7.5 (5H, m).

Elemental analysis for C₁₃ H₂₂ NO₆ P₂ SNa.1/2H₂ O, Calcd.: C, 37.69; H,5.60; N, 3.38 Found: C, 37.31; H, 5.48; N, 3.41

What is claimed is:
 1. A bisphosphonic acid derivative of the generalformula ##STR143## wherein A is a pyridyl group which is unsubstitutedor optionally substituted with 1 to 4 substituents selected from thegroup consisting of (1) a halogen atom, (2) nitro, (3) a C₁₋₇ alkylwhich may be substituted with 1 to 3 substituents selected from thegroup consisting of a halogen atom, hydroxy and a C₁₋₆ alkoxy, (4) aC₃₋₇ cycloalkyl which may be substituted with 1 to 3 substituentsselected from the group consisting of a halogen atom, hydroxy and a C₁₋₆alkoxy, (5) a hydroxyl, (6) a protected hydroxyl group selected from thegroup consisting of C₁₋₆ alkoxy, C₄₋₆ cycloalkoxy, C₂₋₆ alkenyloxy,C₆₋₁₉ aralkyloxy, C₂₋₇ alkanoyloxy and C₆₋₁₄ aryloxy, each of which maybe substituted with 1 to 3 substituents selected from the groupconsisting of a halogen, hydroxyl and a C₁₋₆ alkoxy, (7) thiol and (8) aprotected thiol group selected from the group consisting of a C₁₋₆alkylthio, a C₄₋₇ cycloalkylthio, a C₇₋₁₉ aralkylthio and a C₂₋₇alkanolythio, each of which may be substituted with 1 to 3 substituentsselected from the group consisting of a halogen, hydroxyl and a C₁₋₆alkoxy; R¹ is hydrogen atom or a lower alkanoyl group; R², R³, R⁴ and R⁵are the same or different and are hydrogen atom or a lower alkyl group;m is 0, 1 or 2; and n is an integer from 2 to 10, or a salt thereof. 2.A compound according to claim 1, wherein the lower alkanoyl representedby R¹ is C₁₋₆ alkyl-carbonyl group.
 3. A compound according to claim 1,wherein the lower alkyl represented by R², R³, R⁴ and R⁵ is C₁₋₄ alkyl.4. A compound which is 4-(2-pyridylthio)butylaminomethylenebisphosphonicacid, or its salt or C₁₋₄ alkyl ester.
 5. A compound which is4-(2-pyridylthio)butylaminomethylenebisphosphonic acid.
 6. A boneresorption inhibitory composition which comprises an effective amount ofa compound of the general formula (I) or a salt thereof according toclaim 1, and a pharmaceutically acceptable carrier, diluent orexcipient.
 7. A method of inhibiting bone resorption comprisingadministering an effective amount of a compound of the formula (I) or asalt thereof according to claim 1, optionally together with apharmaceutically acceptable carrier, diluent or excipient, to a patientsuffering from osteoporosis.